The objective of this study was to determine the clinical pregnancy rate with 2 inseminations compared to a single intrauterine insemination (IUI) in a given cycle using frozen-thawed donor sperm. This was a retrospective study at a university practice; patients were women using donor sperm. We conducted a comparison of single IUI, intracervical insemination (ICI) followed by an IUI on the next day, and double IUI (2 consecutive days); clinical pregnancy rate was the main outcome measure. The cyclespecific and total pregnancy rates were not significantly different between the 3 protocol groups (306 cycles). The average pregnancy rate over 3 cycles was 10.2% for IUI, 15.3% for ICI/IUI, and 13.7% for IUI/IUI (P 5 .47). After controlling for repeated measures per subject and age, gravidity, and use of Clomid, there was no significant difference between protocols. The ICI/IUI (odds ratio [OR] 5 1.70; 95% confidence interval [CI], 0.83-3.51) and IUI/IUI (OR 5 1.5; 95% CI, 0.52-4.33) protocols appeared more likely to result in a clinical pregnancy than the single IUI protocol. Current information on the optimal number of inseminations per cycle using donor sperm is limited. Our large study using 3 protocols found an increase in pregnancy rate with the addition of either an ICI or IUI to a single IUI protocol in a natural or Clomid cycle but did not meet statistical significance. Additional prospective studies are needed to better counsel patients using donor sperm.Key words: Donor sperm, intrauterine inseminations, intracervical inseminations, pregnancy rate, Clomid.J Androl 2012;33:375-380 D onor sperm insemination is the first line of treatment for couples with an azoospermic male partner, single women, and same-sex female couples. All donor sperm samples are cryopreserved, allowing for appropriate testing of the donors for infectious diseases. Intrauterine insemination (IUI) or intracervical insemination (ICI) of the frozen-thawed samples can then be performed to achieve pregnancy. The ICI procedure is less invasive, typically using unwashed sperm, and costs less compared to the IUI procedure. A meta-analysis of 7 prospective randomized studies comparing single ICI or IUI using frozen-thawed donor sperm showed that IUI had a significantly higher monthly fecundity rate (odds ratio [OR] 5 2.4; 95% confidence interval [CI], 1.5-3.8; Goldberg et al, 1999). These studies included both natural cycles and treated (clomiphene or injectable gonadotropins) cycles. More recently, a Cochrane systematic review reported significantly higher cumulative pregnancy rates (OR 5 3.37; 95% CI, 1.9-5.96) and live birth (OR 5 1.98; 95% CI, 1.02-3.86) after 6 treated cycles of frozen-thawed donor sperm IUI compared to ICI (Besselink et al, 2008).